Cited by 644Genomics and Chromatin DynamicsNuclear Structure and FunctionChromosomal and Genetic Variations

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Identification of a nonhistone chromosomal protein associated with heterochromatin in Drosophila melanogaster and its gene.

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James, T C, Elgin, S C·1986·Molecular and Cellular Biology

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Abstract

Monoclonal antibodies were prepared against a fraction of nuclear proteins of Drosophila melanogaster identified as tightly binding to DNA. Four of these antibodies were directed against a 19-kilodalton nuclear protein; immunofluorescence staining of the polytene chromosomes localized the antigen to the alpha, beta, and intercalary heterochromatic regions. Screening of a lambda gt11 cDNA expression library with one of the monoclonal antibodies identified a recombinant DNA phage clone that produced a fusion protein immunologically similar to the heterochromatin-associated protein. Polyclonal sera directed against the bacterial lacZ fusion protein recognized the same nuclear protein on Western blots. A full-length cDNA clone was isolated from a lambda gt10 library, and its DNA sequence was obtained. Analysis of the open reading frame revealed an 18,101-dalton protein encoded by this cDNA. Two overlapping genomic DNA clones were isolated from a Charon 4 library of D. melanogaster with the cDNA clone, and a restriction map was obtained. In situ hybridization with these probes indicated that the gene maps to a single chromosome location at 29A on the 2L chromosome. This general strategy should be effective for cloning the genes and identifying the genetic loci of chromosomal proteins which cannot be readily assayed by other means.

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Authors: James, T. C., & Elgin, S. C.
ORCID: James, T C
Elgin, S C
Venue: Molecular and Cellular Biology
Publisher: Informa UK Limited
Published: 1986-11
Volume: 6
Issue: 11
Pages: 3862-3872
ISSN: 0270-7306 (ISSN-L)
1098-5549
DOI: 10.1128/mcb.6.11.3862
PMC: PMC367149
OpenAlex: W2101092089
Cited by: 643
References: 58
Citations by year: 2026: 2 · 2025: 10 · 2024: 21 · 2023: 7 · 2022: 13 · 2021: 37 · 2020: 16 · 2019: 13 · 2018: 12 · 2017: 10 · 2016: 12 · 2015: 21 · 2014: 26 · 2013: 14 · 2012: 14
FWCI: 2.74
Type: journal article
Language: en
Open access: green
www.ncbi.nlm.nih.gov
Locations: Molecular and Cellular Biology (publishedVersion)
PubMed Central (publishedVersion)
PubMed (publishedVersion)
Funders: American Cancer Society
National Institutes of Health
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Nuclear Structure and Function
Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Chromosomal and Genetic Variations
Keywords: non-histone proteins, heterochromatin, Drosophila melanogaster, monoclonal antibodies, polytene chromosomes, cDNA cloning, gene mapping, chromosome 2L, chromatin proteins
MeSH terms: Amino Acid Sequence, Animals, Antibodies, Base Sequence, Chromosomal Proteins, Non-Histone, Chromosomes, DNA, Recombinant, Drosophila melanogaster, Electrophoresis, Polyacrylamide Gel, Fluorescent Antibody Technique, Genes, Heterochromatin

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Identification of a nonhistone chromosomal protein associated with heterochromatin in Drosophila melanogaster and its gene.

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